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Rong Lu, Zhixin Tang, Cong Nie, Ying Chen, Yadan Quan; Can Human Retinoblastoma Cell Lines Replacing Primary Cultured Retinoblastoma cells?. Invest. Ophthalmol. Vis. Sci. 2018;59(9):1625.
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© ARVO (1962-2015); The Authors (2016-present)
Retinoblastoma is the most common malignant intraocular tumor in children. Even after years of research, still little is known about the functional properties of RB cells. Y-79 and WERI-RB1 are two widely applied cell lines. However, after many times passaged in vitro, the biological characteristic and behavior of the RB cell line may be changed from the initial status and cannot stand for RB cells in research. We plan to figure out whether a cell line can be a qualified replacement for primary RB cells.
In the present study, we characterized the primary RB cells and 2 retinoblastoma cell lines Y-79 and WERI-Rb1 with regarding to (1) their RB1 mutation status, (2) biological phenotype, (3) growth kinetics, (4) colony formation potential and (5) tumorigenetic ability. The Primary RB cells were cultured from surgical tissue of RB patients at Zhongshan Ophthalmic Center, Sun Yat-Sen University.
As that we hypothesized, part of the cell function of RB cell lines were different from primary cultured RB cells. The biomarkers expression (CD133, nestin, OCT4, GFAP, MAP2 and recoverin) of primary RB cells was more representative than that of cell line with than that of Y-79 or WERI-RB1. Morphologically, primary RB cells, Y-79 and WERI-Rb1 were similar in serum-containing medium. However, in serum-free medium, primary RB cells could form cell-colony which were solid and round cell spheres with cells fused together and difficult to distinguish them as individual cells. While whether Y-79 or Weri-Rb1 cannot form cell-colony in serum-free medium. The Growth rates of these 3 types of RB cells are different. Y-79 was one of the fastest growing cell lines with a doubling time of 33 hours. The primary RB cells grew much slower of above 4.5 days to double their cell numbers. The tumor formation of RB was assayed by subretinal injections of suspension primary cultured RB cells or RB cell lines (Y-79 or WERI-RB1) into the eyes of immunodeficient mice. All 3 types of RB cell injection could produce orthotopic RB mode.
It indicated that part of the biological characteristics of Y79 and WERI-RB1 were changed from primary cultured RB cells, including bio-phenotype and colony formation potential. While Y-79 and WERI-RB1 cell lines could be a good substitute for primary RB cells to effectively produce orthotopic RB mode. It is important to choose the right type of RB cell for a certain research.
This is an abstract that was submitted for the 2018 ARVO Annual Meeting, held in Honolulu, Hawaii, April 29 - May 3, 2018.
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